Abstract
Since their discovery 25 years ago mechanosensitive (MS) ion channels have been the topic of intensive scientific research. Much of what we know about structure and function of these channels derives from prokaryotic sources, and from patch clamp electrophysiology techniques and X-ray crystallography. But as technology develops so too do the methods employed to investigate the structure and function of these channels become more diverse. Techniques such as electron paramagnetic resonance (EPR) spectroscopy, Förster resonance energy transfer (FRET) imaging and nuclear magnetic resonance (NMR) spectroscopy have all been used to increase our understandings of these channels. But with every new technique come new challenges in sample preparation. Just what is the best way to prepare a MS ion channel for FRET imaging? Which mutants need to be generated? Can we incorporate a MS channel protein into an artificial lipid or will we have to perform the experiments in vivo? To help answer these questions we have documented some of the most common techniques that have been used to investigate MS ion channels to date. We discuss what these techniques have been able to tell us about MS ion channel structure, their molecular dynamics, and just how these channels are capable of responding to mechanical forces exerted from their surrounding lipid bilayer.
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The work from our laboratory discussed in the manuscript was supported by the Australian Research Council (DP0769983), the National Health & Medical Research Council of Australia (635525), and the Yamada Science Foundation.
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Cranfield, C. et al. (2012). Force from Lipids: A Multidisciplinary Approach to Study Bacterial Mechanosensitive Ion Channels. In: Kamkin, A., Lozinsky, I. (eds) Mechanically Gated Channels and their Regulation. Mechanosensitivity in Cells and Tissues, vol 6. Springer, Dordrecht. https://doi.org/10.1007/978-94-007-5073-9_1
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